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Background NASH-related hepatocellular carcinoma (NASH-HCC) arises in a fibrotic, lipid-rich liver microenvironment where extracellular matrix (ECM) stiffening activates mechanosensitive pathways (e.g., Piezo1) that favor immunosuppressive macrophage polarization. In parallel, dysregulated copper metabolism promotes oxidative stress, ECM remodeling, and resistance to cuproptosis. We aim to map how Notch1 links copper dysregulation with mechanical signaling, using a stiffness-tunable 3D bio-printed liver model to study the NASH-to-HCC transition.Methods Notch paralogs were profiled by transcript and immunohistochemical analyses in clinical cohorts, with survival association tested by Cox regression. Fibrosis severity (Sirius Red) and correlations among Notch1, ATP7B (copper export), Piezo1 (mechano-sensing), and M2 macrophage markers (CD163/CD206) were assessed using local cohorts and public datasets. Cuproptosis-related gene programs were analyzed by WGCNA and GO/KEGG enrichment. Functional validation used γ-secretase inhibitor (GSI) treatment in HepG2 cells. A fibrosis-stiffness AlgMA hydrogels and a coaxial 3D-printed liver model was built to mimic the soft NASH and stiff NASH-HCC microenvironments.Results Notch1 was the dominant Notch paralog upregulated in NASH-HCC and associated with worse overall and disease-free survival, remaining an independent prognostic factor with UICC grade ( IDDF2026-ABS-0452 Figure 1(A-C)). Notch1 positively correlated with fibrosis severity and with ATP7B and Piezo1 (IDDF2026-ABS-0452 Figure 2(A)). Piezo1 was elevated in NASH-HCC, predicted poorer survival, and co-localized with CD206+ macrophages in stiff regions (IDDF2026-ABS-0452 Figure 2(B-D)). Cuproptosis-related genes were enriched in immune regulation and collagen-related pathways (IDDF2026-ABS-0452 Figure 3(A,B)). GSI blocked Notch1/Hes1 signaling and produced dose-dependent reductions in ATP7B and Piezo1 (IDDF2026-ABS-0452 Figure 3(C,D)). The AlgMA and coaxial bioprinted models reproduced fibrosis-range stiffness and increased lipid staining in the stiff NASH-HCC compartment (IDDF2026-ABS-0452 Figure 4(A-C)).Conclusions Notch1 integrated copper-metabolic and mechanical pathways to promote ECM remodeling and M2-like macrophage reprogramming, facilitating NASH-HCC progression. Collectively, our 3D bio-printed liver disease platform enabled mechanistic studies and therapeutic screening of this pathway.Abstract IDDF2026-ABS-0452 Figure 1Abstract IDDF2026-ABS-0452 Figure 2Abstract IDDF2026-ABS-0452 Figure 3Abstract IDDF2026-ABS-0452 Figure 4